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Kidney Week

Abstract: SU-OR03

Incorporation of Urine-Derived Stem Cells into Kidney Organoids Derived from Human Induced Pluripotent Stem Cells

Session Information

Category: Acute Kidney Injury

  • 103 AKI: Mechanisms

Authors

  • Bejoy, Julie, Vanderbilt University Medical Center, Nashville, Tennessee, United States
  • Welch, Richard C., Vanderbilt University Medical Center, Nashville, Tennessee, United States
  • Woodard, Lauren Elizabeth, Vanderbilt University Medical Center, Nashville, Tennessee, United States

Group or Team Name

  • Woodard Lab
Background

Donor-derived somatic cells or stem cells can be differentiated into renal cell types for disease modeling, drug screening, or therapeutic studies. Modeling of kidney disease with kidney organoids derived from human induced pluripotent stem cells (hiPSCs) has been termed a “kidney in a dish.” The recent advances in stem cell-based therapies have shown great promise for the treatment of kidney injuries. To evaluate the therapeutic properties, we studied the incorporation of urine-derived stem cells (USCs) into a kidney organoid model of acute kidney injury. UCSs are viable cells from urine which can be expanded in vitro for more than ten passages. There is evidence suggesting that USCs are most likely cultured glomerular parietal epithelial cells.

Methods

For this project, we cultured kidney organoids from fibroblast-derived hiPSCs by the established protocol from Takasato and Little, following optimization.Co-culturing of USCs labeled with a membrane dye and Day 25 kidney organoids revealed that USCs incorporated into the organoids efficiently within two days of the co-culture. For injury models, we established nephrotoxicity in the proximal tubule by adding the nephrotoxic drug Cisplatin (5 μm) at Day 21 of kidney organoid culture.

Results

The kidney organoids derived form IPSCs expressed the kidney cell type markers ECAD (distal tubule), GATA3 (collecting duct), LTL (proximal tubule) and NEPHRIN (Glomeruli) at Day 21.The organoids were then treated with 5 x 104 USCs at Day 22 for 48 hours and evaluated for the expression of kidney injury molecule-1 (KIM-1). Immunostaining revealed that KIM-1 expression was significantly reduced in the organoids treated with USCs compared to the organoids without USCs, suggesting a positive therapeutic impact of USCs. We are currently performing RNAseq on three sets of whole kidney organoids (Control, +Cisplatin, +Cisplatin+USC) to provide detailed interrogation of cellular apoptosis and related signaling pathways in these three different sets of organoids.

Conclusion

The ability of USCs to reduce KIM-1 expression in human kidney organoids suggests that further investigation into the therapeutic potential of USC for treatment of acute kidney injury is warranted.

Funding

  • Private Foundation Support